RL-65 (ATCC® CRL-10354)

Organism: Rattus norvegicus, rat  /  Cell Type: Squamous Epithelium,Epithelial  /  Tissue: lung  / 

Permits and Restrictions

View Permits

Organism Rattus norvegicus, rat
Tissue lung
Cell Type Squamous Epithelium,Epithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Age 5 days
Strain Sprague-Dawley
Storage Conditions liquid nitrogen vapor phase
Genes Expressed
cytoskeletal proteins (alpha keratin, actin, desmin, vimentin, tubulin); fibronectin; laminin
Cellular Products
cytoskeletal proteins (alpha keratin, actin, desmin, vimentin, tubulin); fibronectin; laminin
Tumorigenic No
Effects
No, in immunosuppressed mice
Yes, in semisolid medium.
Comments

The cells are maintained in a serum free medium. If grown in media containing serum, the properties of the cells will change.

The cells exhibit different characteristic when grown with or without retinoic acid.

In the presence of retinoic acid (50 nM), the cells resemble low non-keratinized or squamous epithelium with densely packed colonies. In the absence of retinoic acid, the cells form a keratinized epithelium.

Long term cultures in the absence of retinoic acid form a stratified highly keratinized epithelium with large networks of epithelial filaments.

Complete Growth Medium A 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium with 2.5 mM L-glutamine adjusted to contain 1.2 g/L sodium bicarbonate and supplemented with 0.005 mg/ml insulin, 0.01 mg/ml human transferrin, 0.1 mM ethanolamine, 0.1 mM phosphoethanolamine, 25 nM selenium, 500 nM hydrocortisone, 0.005 mM forskolin, and bovine pituitary extract (0.15 mg protein per ml).
Subculturing

Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of culture medium and aspirate cells by gently pipetting.
  5. Transfer the cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 7 minutes.
  6. Resuspend the cell pellet with the recommended complete growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
  7. Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:10 to 1:20 is recommended
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation
Complete culture medium 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Name of Depositor Genentech, Inc.
References

Mather JP, Roberts PE. Method of isolating lung cell line. US Patent 5,364,785 dated Nov 15 1994

Roberts PE, et al. A novel epithelial cell from neonatal rat lung: isolation and differentiated phenotype. Am. J. Physiol. 259: L415-L425, 1990. PubMed: 2260675

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
FAQ's
  1. Medium preparation for CRL-10354


    Date Updated: 2/21/2014

References

Mather JP, Roberts PE. Method of isolating lung cell line. US Patent 5,364,785 dated Nov 15 1994

Roberts PE, et al. A novel epithelial cell from neonatal rat lung: isolation and differentiated phenotype. Am. J. Physiol. 259: L415-L425, 1990. PubMed: 2260675

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.